Allelic exclusion and peripheral reconstitution by TCR transgenic T cells arising from transduced human hematopoietic stem/progenitor cells

Mol Ther. 2013 May;21(5):1044-54. doi: 10.1038/mt.2013.8. Epub 2013 Feb 5.


Transduction and transplantation of human hematopoietic stem/progenitor cells (HSPC) with the genes for a T-cell receptor (TCR) that recognizes a tumor-associated antigen may lead to sustained long-term production of T cells expressing the TCR and confer specific antitumor activity. We evaluated this using a lentiviral vector (CCLc-MND-F5) carrying cDNA for a human TCR specific for an HLA-A*0201-restricted peptide of Melanoma Antigen Recognized by T cells (MART-1). CD34(+) HSPC were transduced with the F5 TCR lentiviral vector or mock transduced and transplanted into neonatal NSG mice or NSG mice transgenic for human HLA-A*0201 (NSG-A2). Human CD8(+) and CD4(+) T cells expressing the human F5 TCR were present in the thymus, spleen, and peripheral blood after 4-5 months. Expression of human HLA-A*0201 in NSG-A2 recipient mice led to significantly increased numbers of human CD8(+) and CD4(+) T cells expressing the F5 TCR, compared with control NSG recipients. Transduction of the human CD34(+) HSPC by the F5 TCR transgene caused a high degree of allelic exclusion, potently suppressing rearrangement of endogenous human TCR-β genes during thymopoiesis. In summary, we demonstrated the feasibility of engineering human HSPC to express a tumor-specific TCR to serve as a long-term source of tumor-targeted mature T cells for immunotherapy of melanoma.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles*
  • Animals
  • Antigens, CD34 / metabolism
  • Blood Cells / cytology
  • Blood Cells / metabolism
  • Bone Marrow Cells / cytology
  • Bone Marrow Cells / metabolism
  • Complementarity Determining Regions / chemistry
  • Complementarity Determining Regions / genetics
  • Epitopes, T-Lymphocyte / immunology
  • Female
  • Gene Order
  • Genetic Vectors / genetics
  • Graft Survival
  • Hematopoietic Stem Cell Transplantation
  • Hematopoietic Stem Cells / metabolism*
  • Humans
  • Interferon-gamma / biosynthesis
  • Lentivirus / genetics
  • Male
  • Melanoma / genetics
  • Melanoma / immunology
  • Melanoma / therapy
  • Mice
  • Receptors, Antigen, T-Cell / genetics*
  • Receptors, Antigen, T-Cell / metabolism
  • Receptors, Antigen, T-Cell, alpha-beta / chemistry
  • Receptors, Antigen, T-Cell, alpha-beta / genetics
  • T-Lymphocyte Subsets / cytology
  • T-Lymphocyte Subsets / immunology
  • T-Lymphocyte Subsets / metabolism*
  • Thymocytes / cytology
  • Thymocytes / metabolism
  • Transduction, Genetic
  • Transplantation, Heterologous


  • Antigens, CD34
  • Complementarity Determining Regions
  • Epitopes, T-Lymphocyte
  • Receptors, Antigen, T-Cell
  • Receptors, Antigen, T-Cell, alpha-beta
  • Interferon-gamma